Daniel:Notebook/HiResChrPaint/2013-4-22: Difference between revisions
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>Djacobse (Created page with "=Probe Prep S3 4/22/2013= Back to Calendar Purpose: Start new probe synthesis (from scratch) to better assess methods. Getting low yields o...") |
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but I think that the reason is I have much lower yields than Noi's original run. Going to try the entire synthesis, focusing on only | but I think that the reason is I have much lower yields than Noi's original run. Going to try the entire synthesis, focusing on only | ||
2 sets, V4S3 and V6S3. This way I can focus my attention better and use a more appropriately sized trial run. | 2 sets, V4S3 and V6S3. This way I can focus my attention better and use a more appropriately sized trial run. | ||
==qPCR | ==Expansion qPCR== | ||
1. Set up 8 reactions for each of the | 1. Set up 8 reactions for each of the 2 probes (16 reactions total) | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |
Revision as of 19:51, 22 April 2013
Probe Prep S3 4/22/2013
Purpose: Start new probe synthesis (from scratch) to better assess methods. Getting low yields on dye coupling reaction, but I think that the reason is I have much lower yields than Noi's original run. Going to try the entire synthesis, focusing on only 2 sets, V4S3 and V6S3. This way I can focus my attention better and use a more appropriately sized trial run.
Expansion qPCR
1. Set up 8 reactions for each of the 2 probes (16 reactions total)
per rxn | V4S1 | V4S2 | V4S3 | V4S4 | |
20 nM OligoPool | 0.2 | 1.7 | 1.7 | 1.7 | 1.7 |
2X Kapa SYBR qPCR MM | 50 | 425 | 425 | 425 | 425 |
100 uM AP1V41U | 0.4 | 3.4 | 3.4 | 3.4 | 3.4 |
100 uM AP2V4 | 0.4 | 3.4 | 3.4 | 3.4 | 3.4 |
2 mM aa-dUTP | 3 | 25.5 | 25.5 | 25.5 | 25.5 |
H20 | 46 | 391 | 391 | 391 | 391 |
Total Volume | 100 | 850 | 850 | 850 | 850 |
per rxn | V6S1 | V6S2 | V6S3 | V6S6 | |
20 nM OligoPool | 0.2 | 1.7 | 1.7 | 1.7 | 1.7 |
2X Kapa SYBR qPCR MM | 50 | 425 | 425 | 425 | 425 |
100 uM AP1V61U | 0.4 | 3.4 | 3.4 | 3.4 | 3.4 |
100 uM AP2V6 | 0.4 | 3.4 | 3.4 | 3.4 | 3.4 |
2 mM aa-dUTP | 3 | 25.5 | 25.5 | 25.5 | 25.5 |
H20 | 46 | 391 | 391 | 391 | 391 |
Total Volume | 100 | 850 | 850 | 850 | 850 |